Rapid and accurate identification of highly infectious and pathogenic disease is extremely important for the early forecast of outbreak risk and the effective deployment of medical and administrative countermeasures. Unfortunately, current diagnostics for use at the point-of-care (POC) are largely immunological assays that often lack adequate sensitivity, specificity, and multiplex capacity for an accurate early diagnosis. Nucleic acid- based diagnostic products developed or under development requires a relatively expensive dedicated instrument that is often prohibitive to small clinics and resource-limited settings and unsuitable for pandemic high capacity demand. We propose to develop an instrumentation-free, self-contained, low cost (<$20/unit), disposable, rapid (<30 minute, sample-to-answer), nucleic acid POC diagnostic device (MTIDx) capable of detecting influenza A, influenza B, RSV and RNaseP internal control (4-plex). The final MTIDx product is designed to be a FDA approved, CLIA-waived (Clinical Laboratory Improvement Amendments of 1988) nucleic acid (NA)-based diagnostic device for analyzing human nasal swab samples. MTIDx (Multiple Target Identification Diagnostic) will be a pocket-sized, device that is easy to operate with minimal user-intervention and also has an option to be powered by a 9V battery for use in resource-limited/field settings. It will be suitable for high volume manufacturing allowing rapid production of millions of units to meet surge capacity during an epidemic outbreak. The diagnostic system is based on the integration of Mesa Tech's proprietary technologies of nucleic acid extraction, rapid amplification, lateral flow-based (LF-based) colorimetric detection components with simple fluid control and microheater solutions into one easily used, low cost, disposable device. In Phase 1 project, we have successfully developed of integrated device through a rapid prototyping approach. Work in this Phase II project will focus on the optimization of Phase I design to include more refined design for a low complexity, CLIA-waved product as well as cost effective and environment friendly reusable electronic circuit component concept that allows the wide and rapid adaption at POC. We will further optimize the development of lyophilized reagents to confer ambient storage compatibility, development of stable buffer reagents and capsulation methods, design for manufacturing optimizations and conduct performance evaluation on clinical samples. The primary Phase II deliverable is the design of MTIDx ready for manufacturing (Design For Manufacturing) that offers a sample-to-answer 4-plex diagnostic test (Flu A, FluB, RSV, and RNase P control) under 30 minutes prior to the follow up manufacturing, FDA and marking effort.